Method of making a vaccine

ABSTRACT

A method of producing attenuated cells for a mammalian vaccine comprising killing or attenuating the cells with a high energy, low heat UV light.

COPYRIGHT NOTICE

A portion of the disclosure of this patent contains material that issubject to copyright protection. The copyright owner has no objection tothe reproduction by anyone of the patent document or the patentdisclosure as it appears in the Patent and Trademark Office patent filesor records, but otherwise reserves all copyright rights whatsoever.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to a method and system for makingvaccines. In particular, it relates to creating attenuated cells forcreating a vaccine in large quantities especially useful for use inindividually produced vaccines. It also is a method for creatingantibodies.

Description of Related Art

The production of a vaccine and its usefulness are well-known. Aninfectious cell is attenuated but not so destroyed that it does notstill create an immune response, producing antibodies without causingthe user to contract the disease. While such an approach does not workwith all infections cells, its usefulness is unquestionable.

One of the biggest problems is the process of killing the cells withoutdamage. Even more problematic is the time it takes to kill the cells,which can be drawn out, leading to long lead times for producing largequantities of a vaccine, especially during outbreaks such as the flu orthe like. One version of the attenuation process is the exposure of Phaemolytica to UV irradiation for 60 minutes using low level UVirradiation. Since temperatures, as taught in the art, need to be around58 degrees F. or less, exposure to high energy UV irradiation is out ofthe question because of the high heat generated. Further, it is notknown if such high energy UV will damage cellular proteins andcarbohydrates necessary to create an immune response. Accordingly, thecurrent UV low energy practice is not widely utilized, if at all.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to the use of high energy, low heat UVsource which rapidly kills cells without damaging structures necessaryfor the production of an antibody response. It is discovered that such aprocess takes mere seconds rather than an hour or more, leading to rapidproduction of vaccines from live cell cultures. Such cells can then beintroduced into patients as vaccines, or to stimulate additionalantibody production in an already infected individual.

Accordingly, in one embodiment, there is a method of producingattenuated infectious cells comprising:

-   -   a) isolating a plurality of infectious cells;    -   b) selecting a high energy UV light source that delivers a low        heat UV containing light to the end of a light guide; and    -   c) exposing the infectious cells to the light from the end of        the light guide at sufficient distance and time to kill the        cells without damaging cellular proteins and carbohydrates to        produce attenuated cells.

In another embodiment, there is a composition comprising a vaccine withattenuated infectious cells made from exposure to a high energy UV lightsource that delivers a low heat UV containing light to the end of alight guide.

DETAILED DESCRIPTION OF THE INVENTION

While this invention is susceptible to embodiment in many differentforms, there is shown in the drawings, if any, and will herein bedescribed in detail specific embodiments, with the understanding thatthe present disclosure of such embodiments is to be considered as anexample of the principles and not intended to limit the invention to thespecific embodiments shown and described. In the description below, likereference numerals, if any, are used to describe the same, similar orcorresponding parts in the several views of the drawings, if any. Thisdetailed description defines the meaning of the terms used herein andspecifically describes embodiments in order for those skilled in the artto practice the invention.

Definitions

The terms “about” and “essentially” mean±10 percent.

The terms “a” or “an”, as used herein, are defined as one or as morethan one. The term “plurality”, as used herein, is defined as two or asmore than two. The term “another”, as used herein, is defined as atleast a second or more. The terms “including” and/or “having”, as usedherein, are defined as comprising (i.e., open language). The term“coupled”, as used herein, is defined as connected, although notnecessarily directly, and not necessarily mechanically.

The term “comprising” is not intended to limit inventions to onlyclaiming the present invention with such comprising language. Anyinvention using the term comprising could be separated into one or moreclaims using “consisting” or “consisting of” claim language and is sointended.

Reference throughout this document to “one embodiment”, “certainembodiments”, and “an embodiment” or similar terms means that aparticular feature, structure, or characteristic described in connectionwith the embodiment is included in at least one embodiment of thepresent invention. Thus, the appearances of such phrases or in variousplaces throughout this specification are not necessarily all referringto the same embodiment. Furthermore, the particular features,structures, or characteristics may be combined in any suitable manner inone or more embodiments without limitation.

The term “or” as used herein is to be interpreted as an inclusive ormeaning any one or any combination. Therefore, “A, B or C” means any ofthe following: “A; B; C; A and B; A and C; B and C; A, B and C”. Anexception to this definition will occur only when a combination ofelements, functions, steps or acts are in some way inherently mutuallyexclusive.

The drawings featured in the figures, if any, are for the purpose ofillustrating certain convenient embodiments of the present invention,and are not to be considered as limitation thereto. The term “means”preceding a present participle of an operation indicates a desiredfunction for which there is one or more embodiments, i.e., one or moremethods, devices, or apparatuses for achieving the desired function andthat one skilled in the art could select from these or their equivalentin view of the disclosure herein and use of the term “means” is notintended to be limiting.

As used herein, the term “attenuated” refers to a virus or bacterium orother pathogen that has been modified to reduce its virulence to theinfection the cell would cause while maintaining the virus or bacteria'sability to cause an immune response. The cell can either be alive ordead. The term “live attenuated vaccine” is one which the cells remainviable for infecting and replicating within a target host. The presentinvention can be used to modify a wide variety or viruses, e.g.,adenovirus, measles, mumps, rubella, influenza, chicken pox, smallpox,polio, rotavirus, yellow fever, chikungunya, hantavirus,cytomegalovirus, dengue, Epstein-Barr virus, hepatitis A, B, C, or E,human papilloma virus, encephalitis, HIV, and rabies, to name a few.Attenuated vaccines can be formulated for use in mammals, e.g., humans.Furthermore, the present invention can be designed, following theteachings herein, to be grown in an avian system for use as a vaccine ina mammalian, vice versa, or using other viral expression systems (e.g.,insect cells) for use in non-insects. Bacteria for use could include,but is not limited to, Staphylococcus aureus, Bacillus athracis,Treponema pallidum, strepococcus, Nesseria meningitis, Escherichia coli,Pseudomonas aeruginosa, tuberculosis, Haemophila influenzae, Enterococcifaecalis, Clostridium difficile, legionella, listeria, salmonella,clostridia, leptospira, borellia, Helicobacter pylori, and the like.

As used herein, the term “vaccine”, “vaccination”, and “vaccinating”refer to compositions and methods for modulating an immune response to aselected antigen such that the response is more efficient, more rapid,greater in magnitude, and/or more easily induced.

As used herein, the term “modulating an immune response” refers to thestimulation and/or activation of an immune response to a selectedantigen, but it also refers to the suppression, elimination, orattenuation of an immune response to a selected antigen.

As used herein, the term “antigen” refers to a molecule that caninitiate a humoral and/or a cellular immune response in a recipient tothe antigen. Antigens can be any type of biologic molecule including,for example, simple intermediary metabolites, sugars, lipids, andhormones as well as macromolecules such as complex carbohydrates,phospholipids, nucleic acids, and proteins. Common categories ofantigens include, but are not limited to, viral antigens, bacterialantigens, fungal antigens, protozoa and other parasitic antigens, tumorantigens, antigens involved in autoimmune disease, allergy and graftrejection, and other miscellaneous antigens.

The compositions, vaccines, or formulations of the invention can beused, for example, to modulate an immune response in a mammal such as ahuman. Vaccination with infectious cell live attenuated vaccines (LAVs)is an effective way for prevention of infectious disease.

As used herein, the term “infectious cells” refers to a whole-cellpathogen with intact DNA.

As used herein, the term “isolating a plurality of infectious cells”refers to whole-cell pathogens either homogenous or heterogenous.

As used herein, the term “high intensity UV light source that delivers alow heat UV light” refers to a device that has a light source producinga spectrum of UV light capable of killing a microorganism, such astaught in US patent publication number US2017/0028089 published on Feb.2, 2017 in the name of Kurt A. Garrett and U.S. patent application Ser.No. 15/712,559 filed on Sep. 22, 2017 in the name of Kurt A. Garrett. Inparticular, it produces a wide UV spectrum (i.e. more than just anisolated wavelength) even though it can produce other spectrums of lightand, in one embodiment, the light produces a high UV output. It consistsof a high intensity light, a dichroic reflector which focuses the lightand removes heat and then delivers the UV light to a light tube wherethe light coming out of the far end of the light tube is used to killcancer cells without damaging underlying physical structure thusminimizing or eliminating damage to healthy cells near the cancer cells.

As used herein, the term “high intensity light” refers to a bulb of anykind which produces a sterilizing UV light. This can be UVA, UVB, UVC,or a combination. Regular bulbs, but also high intensity discharge (HID)bulbs, are also embodiments of the invention. For example, a highintensity mercury xenon (HgXe) bulb can be utilized. These types ofbulbs are high UV output bulbs. In general, the light output of somebulbs of the invention is from about 0.1 J/cm² to about 50.0 J/cm². Italso includes arc type lamps when they are focused properly to the lighttube.

As used herein, the term “high intensity light” refers to light outputof about at least 80 lumens per watt output. In order to achieve thishigh intensity light output, one cannot use low or medium pressure lampsthat produce UV light, as they do not produce enough light output. Inorder to achieve the high intensity output needed, one can add to thearc discharge lamp's light output an elliptical reflector whichcollimates the polychromatic light into still greater intensity(intensity being understood as energy per area) of about 100 lumens perwatt (i.e. producing the high intensity light output needed).

As used herein, the term “dichroic reflector” refers to any of adichroic focus, reflector, mirror, lens, or the like that takes lightfrom the light source and allows some or all of the thermal energy topass through the reflector while taking the light, especially the UVlight, to be reflected for focusing. In one embodiment, there may bemore than one dichroic reflector but at least one must focus the lightto the light pipe. The dichroic reflector can be any shape that works toeither remove heat or focus the light but, in one embodiment, it is anelliptical shape for focusing. In one embodiment, an elliptical dichroicreflector is used with an arc lamp. This is different from a dichroicfilter which only filters or reflects light but does not pass heatwavelengths through it. The dichroic filter can be a powered orunpowered device.

As used herein, the term “polychromatic” refers to light comprisingmultiple wavelengths of light.

As used herein, the term “sufficient distance and time” refers to thetime period and distance from the cells that light produced by thedevice is exposed (light shining on it) to a cell in order to kill it.In one embodiment, it is from about 0.01 seconds to about 5 seconds. Inone embodiment, a shutter is utilized to open, close, and modulate thepassage of light from the light source to the cancer cell. The exposurecan be directly from the end of the light tube or extended via a lightfiber at the end of the light tube, especially for insertion of thefiber into a cell tumor either directly or through the skin of theanimal or human.

Following irradiation, cells are prepared for administration as avaccine by formulation in an effective immunization dosage with apharmaceutically acceptable carrier. An effective immunization dosage isdefined herein as being that amount which will induce complete orpartial immunity (elicit a protective immune response) in a treatedanimal against subsequent challenge with virulent cells. Immunity isconsidered as having been induced in a population of treated mammalswhen the level of protection for the population is significantly higherthan that of an unvaccinated control group. The appropriate effectivedosage can be readily determined by the practitioner skilled in the art.

The cells are prepared for administration by formulation in apharmaceutically acceptable carrier such as physiological saline,mineral oil, vegetable oils, aqueous sodium carboxymethyl cellulose, oraqueous polyvinylpyrrolidone. The vaccine formulations may also containoptional adjuvants, antibacterial agents, or other pharmaceuticallyactive agents as are conventional in the art. Without being limitedthereto, suitable adjuvants include, but are not limited to, mineraloil, vegetable oils, alum, Freund's incomplete adjuvant, and Freund'sincomplete adjuvant with oils being embodiments. Still other embodimentadjuvants include microparticles or beads of biocompatible matrixmaterials. The microparticles may be composed of any biocompatiblematrix materials as are conventional in the art including, but notlimited to, agar and polyacrylate. The practitioner skilled in the artwill recognize that other carriers or adjuvants may be used as well.

In accordance with a preferred embodiment, the cells may be incorporatedinto microparticles or microcapsules to prolong the exposure of theantigenic material to the subject animal and hence protect the animalagainst infection for long periods of time. The microparticles andcapsules may be formed from a variety of well-known inert, biocompatiblematrix materials using techniques conventional in the art. Without beinglimited thereto, suitable matrix materials include natural or syntheticpolymers such as alginates, poly(lactic acid), polylactic/glycolicacid), poly(caprolactone), polycarbonates, polyamides, oxide, andparticularly agar and polyacrylates.

The vaccines of the invention may be administered to the subject mammalintramuscular or transthoracic injection, or by aerosol. However,subcutaneous injection is preferred for practical considerations. Thevaccine may be administered in a single dose or in a plurality of doses.In accordance with a preferred embodiment, the vaccine may beadministered in two doses about 2 to 6 weeks apart, most preferablyabout 2-3 weeks apart. The subject animals may be vaccinated at anytime, although it is preferred to administer the vaccine shortly(optimally about 10 days to two weeks) before periods of anticipatedstress, such as during shipping or other handling. It is also envisionedthat the vaccine may be administered to pregnant animals prior to birthto increase production of hyper-immune colostrum.

Accordingly, in the practice of the invention, selected infectious cellswhich have an antigen or other chemical which illicits an immuneresponse in a mammal is isolated using methods known in the art. Thehigh energy UV light source that delivers a low heat UV containing lightto the end of the light guide is selected, for example, such as shown inUS Patent publication number US2017/0028089 published on Feb. 2, 2017 inthe name of Kurt A. Garrett and U.S. patent application Ser. No.15/712,559 filed on Sep. 22, 2017 in the name of Kurt A. Garrett. Thelight is used to kill or at least render the cell nondeliterious. Sinceexposure times are a matter of seconds rather than minutes, largequantities of cells can be treated in a short period of time. Forexample, a conveyor belt type system could be utilized or the light on arobotic arm could cover large areas of surface in a meticulous manner.

Those skilled in the art to which the present invention pertains maymake modifications resulting in other embodiments employing principlesof the present invention without departing from its spirit orcharacteristics, particularly upon considering the foregoing teachings.Accordingly, the described embodiments are to be considered in allrespects only as illustrative, and not restrictive, and the scope of thepresent invention is, therefore, indicated by the appended claims ratherthan by the foregoing description or drawings, if any. Consequently,while the present invention has been described with reference toparticular embodiments, modifications of structure, sequence, materialsand the like apparent to those skilled in the art still fall within thescope of the invention as claimed by the applicant.

What is claimed is:
 1. A method of producing a live or dead attenuatedinfectious cells comprising: a) isolating a plurality of infectiouscells; b) selecting a high energy UV light source that delivers a lowheat UV containing light to the end of a light guide; c) selecting theUV light source and output so that UV energy from the light source canselectively attenuate cells; and d) exposing the infectious cells to thelight from the end of the light guide at sufficient distance and whereinthe cells are exposed to the light from about 0.1 seconds to about 5seconds to attenuate the cells without damaging cellular proteins andcarbohydrates that produce the immune response to produce attenuatedcells.
 2. The method according to claim 1 wherein the attenuated cellsare formulated into a vaccine.
 3. The method according to claim 2wherein the vaccine is administered to a mammal in a dose effective toelicit an immune response that creates antibodies to the infectiouscells.
 4. A method of producing a live or dead attenuated infectiouscells comprising: a) isolating a plurality of infectious cells; b)selecting a high energy UV light source having an output of at leastabout 80 lumens per watt that delivers a low heat UV containing light tothe end of a light guide; c) selecting the UV light source and output sothat UV energy from the light source can selectively attenuate cells;and d) exposing the infectious cells to the light from the end of thelight guide at sufficient distance and time to attenuate the cellswithout damaging cellular proteins and carbohydrates that produce theimmune response to produce attenuated cells.
 5. The method according toclaim 4 wherein the attenuated cells are formulated into a vaccine. 6.The method according to claim 5 wherein the vaccine is administered to amammal in a dose effective to elicit an immune response that createsantibodies to the infectious cells.
 7. The method according to claim 1wherein the cells are exposed to the light from about 0.1 seconds toabout 5 seconds.